Human CD95 ELISA Assay


The Human CD95 ELISA Assay is to be used for the in-vitro quantitative determination of soluble CD95 (APO-1, Fas) in human serum, plasma, buffered solutions or cell culture medium. The assay will recognize both natural and recombinant human sCD95. The Eagle Biosciences Human CD95 ELISA Assay kit is for Research Use Only. 

SKU: C9531-K01 Categories: , ,

Human CD95 ELISA Assay

The Human CD95 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 47 pg/ml
Dynamic Range: 93.75 – 3000 pg/ml
Incubation Time: 3h 45min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: FAS, APO-1

Assay Background

Fas (APO-1, CD95) is a transmembrane protein belonging to the TNF receptor family, capable of mediating apoptotic death of transformed cells as well as activated human T cells. The cell death signal can be induced by anti-Fas antibodies or by the Fas (Fas-L) ligand belonging to the TNF family. The in vivo expression of these two Fas and Fas-L molecules is differently regulated since the expression of Fas is pleiotropic whereas Fas-L is rather expressed in immunity sites (testes, ovaries, brain, eye) and by activated T cells. Tumor cells regardless of their tissue origin may also express Fas-L (pathological).

Genetic and immunological studies have shown that the Fas system is on the one hand involved in the process of peripheral clonal deletion allowing the maintenance of self-tolerance, on the other hand that it regulates the amplitude and duration of immune responses. and finally that it represents one of the mechanisms of lysis of the target cells in the antiviral cytotoxic T response.

When Fas-dependent apoptosis can not occur, due to genetic mutations in the Fas or Fas-L genes, a lymphoproliferative syndrome and autoimmune manifestations are observed in humans and mice. Conversely, an exacerbation of the Fas system may be responsible for tissue destruction. For example, the single mouse injection of anti-Fas antibodies induces massive liver destruction and mouse death within a few hours, leading to the hypothesis of Fas involvement in fulminant hepatitis.

Related Products

Human CD178 ELISA Assay Kit
Human TRAIL / Apo2L ELISA Assay Kit
Human CD14 ELISA Assay Kit

Additional Information

Assay Principle

The sCD95 Kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for sCD95 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known sCD95 concentrations, and unknowns are pipetted into these wells.
During the first incubation, the sCD95 antigen and a biotinylated monoclonal antibody specific for sCD95 are simultaneously incubated.

After washing, the enzyme ( streptavidin-peroxydase ) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution which is acting on the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of sCD95 present in the samples.

  1. Add 100µl of each standard, sample, control and zero (standard diluents) in duplicate to appropriate number of wells
  2. Add 50µl of diluted biotinylated anti-sCD95 to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  4. Remove the cover and wash the plate
  5. Add 100μl of Streptavidin-HRP solution into all wells
  6. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  7. Repeat wash step 4.
  8. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  9. Incubate in the dark for 20-30 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  10. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve

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